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Single-walled carbon nanohorn (SWNH) aggregates inhibited proliferation of human liver cell lines and promoted apoptosis, especially for hepatoma cell lines.

Zhang J, Sun Q, Bo J, Huang R, Zhang M, Xia Z, Ju L, Xiang G - Int J Nanomedicine (2014)

Bottom Line: However, the interactions between the material itself and cancerous or normal cells have seldom been studied.To address this problem, the effects of raw SWNH material on the biological functions of human liver cell lines were studied.Further research on the mechanisms and application in treatment of hepatocellular carcinoma with SWNHs is needed.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, People's Republic of China.

ABSTRACT
Single-walled carbon nanohorns (SWNHs) may be useful as carriers for anticancer drugs due to their particular structure. However, the interactions between the material itself and cancerous or normal cells have seldom been studied. To address this problem, the effects of raw SWNH material on the biological functions of human liver cell lines were studied. Our results showed that unmodified SWNHs inhibited mitotic entry, growth, and proliferation of human liver cell lines and promoted their apoptosis, especially in hepatoma cell lines. Individual spherical SWNH particles were found inside the nuclei of human hepatoma HepG2 cells and the lysosomes of normal human liver L02 cells, implying that SWNH particles could penetrate into human liver cells_and the different interacted mechanisms on human normal cell lines compared to hepatoma cell lines. Further research on the mechanisms and application in treatment of hepatocellular carcinoma with SWNHs is needed.

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TEM images of liver cells.Notes: L02 cells (3×105) and HepG2 cells (3×105) were seeded onto 60 mm noncoated and SWNHs40-coated (0.85 μg/cm2) dishes, respectively, and cultured for 48 hours. The cells were collected and fixed with 3% glutaraldehyde. For TEM, ultra-thin cell slices of 100 nm thickness were cut using an ultramicrotome and mounted on grids. The slices were contrasted with aqueous solution of uranyl acetate and lead citrate, and examined with a JEM-1400 transmission electron microscope (JEOL Ltd, Tokyo, Japan) with accelerating voltage of 80 kV. (A) L02 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (B) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (C) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel B). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside lysosomes of L02 cells. (D) HepG2 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (E) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (F) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel E). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside nuclei of HepG2 cells. (G) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (H) HepG2 cells cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm.Abbreviations: TEM, transmission electron microscope; L02, normal liver cell line; HepG2, human hepatoma cell line; SWNHs40, 0.85 μg of single-walled carbon nanohorns per cm2, or 24 μg per 60 mm dish; SWNH, single-walled carbon nanohorn.
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f8-ijn-9-759: TEM images of liver cells.Notes: L02 cells (3×105) and HepG2 cells (3×105) were seeded onto 60 mm noncoated and SWNHs40-coated (0.85 μg/cm2) dishes, respectively, and cultured for 48 hours. The cells were collected and fixed with 3% glutaraldehyde. For TEM, ultra-thin cell slices of 100 nm thickness were cut using an ultramicrotome and mounted on grids. The slices were contrasted with aqueous solution of uranyl acetate and lead citrate, and examined with a JEM-1400 transmission electron microscope (JEOL Ltd, Tokyo, Japan) with accelerating voltage of 80 kV. (A) L02 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (B) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (C) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel B). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside lysosomes of L02 cells. (D) HepG2 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (E) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (F) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel E). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside nuclei of HepG2 cells. (G) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (H) HepG2 cells cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm.Abbreviations: TEM, transmission electron microscope; L02, normal liver cell line; HepG2, human hepatoma cell line; SWNHs40, 0.85 μg of single-walled carbon nanohorns per cm2, or 24 μg per 60 mm dish; SWNH, single-walled carbon nanohorn.

Mentions: Liver cells were cultured in dishes treated with SWNHs40 (0.85 μg/cm2) for 48 hours and then were collected for TEM measurement. SWNHs could be observed in cells by TEM. SWNHs aggregate was smaller than 100 nm, and localized at lysosomes of L02 (Figure 8B and C) and nuclei of HepG2 cells (Figure 8E and F), respectively. The untreated cells are shown in Figure 8A (L02) and Figure 8D (HepG2). The individual SWNH particles were easily distinguished from their neighbor molecules because of higher Л electronic densities on the surface of SWNHs than those of other molecules in organelles.


Single-walled carbon nanohorn (SWNH) aggregates inhibited proliferation of human liver cell lines and promoted apoptosis, especially for hepatoma cell lines.

Zhang J, Sun Q, Bo J, Huang R, Zhang M, Xia Z, Ju L, Xiang G - Int J Nanomedicine (2014)

TEM images of liver cells.Notes: L02 cells (3×105) and HepG2 cells (3×105) were seeded onto 60 mm noncoated and SWNHs40-coated (0.85 μg/cm2) dishes, respectively, and cultured for 48 hours. The cells were collected and fixed with 3% glutaraldehyde. For TEM, ultra-thin cell slices of 100 nm thickness were cut using an ultramicrotome and mounted on grids. The slices were contrasted with aqueous solution of uranyl acetate and lead citrate, and examined with a JEM-1400 transmission electron microscope (JEOL Ltd, Tokyo, Japan) with accelerating voltage of 80 kV. (A) L02 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (B) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (C) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel B). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside lysosomes of L02 cells. (D) HepG2 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (E) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (F) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel E). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside nuclei of HepG2 cells. (G) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (H) HepG2 cells cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm.Abbreviations: TEM, transmission electron microscope; L02, normal liver cell line; HepG2, human hepatoma cell line; SWNHs40, 0.85 μg of single-walled carbon nanohorns per cm2, or 24 μg per 60 mm dish; SWNH, single-walled carbon nanohorn.
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f8-ijn-9-759: TEM images of liver cells.Notes: L02 cells (3×105) and HepG2 cells (3×105) were seeded onto 60 mm noncoated and SWNHs40-coated (0.85 μg/cm2) dishes, respectively, and cultured for 48 hours. The cells were collected and fixed with 3% glutaraldehyde. For TEM, ultra-thin cell slices of 100 nm thickness were cut using an ultramicrotome and mounted on grids. The slices were contrasted with aqueous solution of uranyl acetate and lead citrate, and examined with a JEM-1400 transmission electron microscope (JEOL Ltd, Tokyo, Japan) with accelerating voltage of 80 kV. (A) L02 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (B) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (C) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel B). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside lysosomes of L02 cells. (D) HepG2 not treated with SWNHs, as control (15,000×). Scale bar represents 1 μm. (E) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (F) HepG2 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (80,000×). Scale bar represents 100 nm (image was enlarged from the rectangular inset of panel E). The arrow shows that there were individual spherical SWNH particles with diameters less than 100 nm inside nuclei of HepG2 cells. (G) L02 cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm. (H) HepG2 cells cultured onto SWNHs40-coated dishes (0.85 μg/cm2) for 48 hours (15,000×). Scale bar represents 1 μm.Abbreviations: TEM, transmission electron microscope; L02, normal liver cell line; HepG2, human hepatoma cell line; SWNHs40, 0.85 μg of single-walled carbon nanohorns per cm2, or 24 μg per 60 mm dish; SWNH, single-walled carbon nanohorn.
Mentions: Liver cells were cultured in dishes treated with SWNHs40 (0.85 μg/cm2) for 48 hours and then were collected for TEM measurement. SWNHs could be observed in cells by TEM. SWNHs aggregate was smaller than 100 nm, and localized at lysosomes of L02 (Figure 8B and C) and nuclei of HepG2 cells (Figure 8E and F), respectively. The untreated cells are shown in Figure 8A (L02) and Figure 8D (HepG2). The individual SWNH particles were easily distinguished from their neighbor molecules because of higher Л electronic densities on the surface of SWNHs than those of other molecules in organelles.

Bottom Line: However, the interactions between the material itself and cancerous or normal cells have seldom been studied.To address this problem, the effects of raw SWNH material on the biological functions of human liver cell lines were studied.Further research on the mechanisms and application in treatment of hepatocellular carcinoma with SWNHs is needed.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, People's Republic of China.

ABSTRACT
Single-walled carbon nanohorns (SWNHs) may be useful as carriers for anticancer drugs due to their particular structure. However, the interactions between the material itself and cancerous or normal cells have seldom been studied. To address this problem, the effects of raw SWNH material on the biological functions of human liver cell lines were studied. Our results showed that unmodified SWNHs inhibited mitotic entry, growth, and proliferation of human liver cell lines and promoted their apoptosis, especially in hepatoma cell lines. Individual spherical SWNH particles were found inside the nuclei of human hepatoma HepG2 cells and the lysosomes of normal human liver L02 cells, implying that SWNH particles could penetrate into human liver cells_and the different interacted mechanisms on human normal cell lines compared to hepatoma cell lines. Further research on the mechanisms and application in treatment of hepatocellular carcinoma with SWNHs is needed.

Show MeSH
Related in: MedlinePlus